Standard Decapping Protocol (NEB #M0608)

1. Set up the following reaction on ice.
   
   

   COMPONENT	20 µl REACTION	FINAL CONC. OR AMOUNT
   mRNA Decapping Enzyme Reaction Buffer (10X)	2 µl	1X
   Total RNA	up to 5 µg
   mRNA Decapping Enzyme	1 µl	5 units/µl
   Nuclease-free water	to 20 µl

2. Gently mix the reaction by pipetting up and down and microfuge briefly.
   
   
3. Incubate at 37°C for 30 minutes.
   
   

Note: The enzyme can be inactivated with EDTA or SDS.

• Use a final concentration of 25 mM EDTA to stop the reaction.
  
  
• Use a final concentration of 2% SDS to stop the reaction.

The reaction volume can be scaled as needed. The incubation time can be increased to 1 hour if necessary.